| The pathogenesis of Ulcerative colitis (UC) was related with many factors, the main of which are immune, genetic, infection and environmental factors, the immune factors which play an important role in. The UC pathogenesis is not clear, the study of its mechanism is mainly focused on immune molecules and inflammatory mediators and other fields, especially the NF-κB pathway, is gradually revealed that the pathogenesis of UC. By acting on the pathway of IλB α, IKK β key molecules, and inhibition of nuclear factor-κB(NF-κB) activation, inhibition of UC intestinal immune and inflammatory responses has become the research focus of UC. The study also found that cyclooxygenase-2(COX-2), inducible nitric oxide synthase(iNOS), myeloperoxidase(MPO), interleukin10(IL-10) and NF-κB pathway is closely related to. COX-2, iNOS, MPO in UC intestinal inflammation in role of Promotion, and IL-10in UC intestinal inflammation in role of inhibition. Chinese medicine is effective in the treatment of UC, and there is no lack of western medicine side effects and high recurrence rate, but its mechanism of action to carry out less, limits the improvement of the level of medical treatment. On the basis of the pre Kuijieling on UC colonic mucosa of rats of NF-κB activation was significantly inhibited, this study further observation of the key elements of UC rat model of NF-κB pathway, IκB and IKKβ as well as closely related to the pathway COX-2, iNOS and of IL-10, etc., and to explore the Kuijieling treatment of UC and the role of deep-level mechanism, and to lay the foundation for the development of high-quality anti-UC Traditional Chinese Drug.1Effect of KD on I κ B α, IKK β prote in i n Co I on i c Mucosa of UC Rats Mode IIκB a, IKK β protein is NF-κB pathways important member, in the NF-kB activation process plays an important role in. This study uses Kui jieling three dose (18.3g/kg,9.2g/kg,4.6g/kg) on trinitrobenzene sulfonic acid (TNBS) method in UC rat model for treatment, after treatment for colonic tissue fixation, preparing pathological sections, immunohistochemical staining, microscopic counting positive cell percentage, and carries on statistics analysis. The results show that the model control group(MC) of IkB a Protein positive rate was47.86±16.52%, significantly lower than the normal control group(NC)82.91±28.00%(P<0.01); Kuijieling in medium dose (KMD) group, Kuijieling in low dose (KLD) group, IκB a Protein expression rate was70.57±21.26%,69.37±25.23%, obviously higher than those in the MC group (P<0.05). In addition the MC group of the IKK β protein positive rate was69.15±19.45%, significantly higher than that in NC group44.74±16.32%(P<0.05); KMD group IKK β protein positive rate was49.79±18.52%, significantly higher than the MC group (P<0.05). Tips of KD could obviously increase the TNBS method UC rat model of colonic mucosa in the IκBa Protein positive cells express rate, reduce the positive expression rate of UC rat model of colon mucosa IKKβ protein.2Effect of KD on COX-2protein in Colonic Mucosa of UC Rats ModelNormal colonic mucosa tissue COX-2expression in inflammatory, epithelial cells and colonic lamina propria mononuclear inflammatory cells, expression, expression of COX-2and UC severity was positively related to. Studies have shown that the expression of COX-2can regulate NF-κB pathway. The experimental group and with the treatment, after the end of treatment of colonic mucosal specimens collection and extraction of whole cell protein, the protein immunoblot (Western blot) method for the detection of colonic mucosal COX-2protein expression levels, with β-actin as reference, to target protein and β-actin density ratio as objective relative protein content, compared analysis. The results show that the (MC) group of colonic mucosa in COX-2protein relative expression capacity of1.189±0.498, significantly higher than that in NC group0.639±0.334(P<0.05); Kuijieling high(KHD), medium, low dose group of colonic mucosa in COX-2protein relative expressions were0.651±0.428,0.643±0.426,0.599±0.340, significantly lower than that in MC group1.189±0.498(P<0.05).Tips of KD on UC in colonic mucosa of rats with COX-2protein expression in inhibition.3Effect of KD on iNOS, MPO activity in Colonic Mucosa of UC Rats Model Nitric oxide synthase (NOS) is a key enzyme of the synthesis of NO, NO synthesis excess can promote the inflammatory response. Inducible nitric oxide synthase (iNOS) is one of the isoenzyme of the NOS, the enzyme involved in the synthesis of NO release, long duration. At present, many studies have shown that iNOS and UC incidence is closely related to the UC when the NO generated by iNOS. iNOS in the level of gene transcription by NF-κB regulation. MPO is an important peroxidase, the increase of MPO content in neutrophils can react in an organization increases, the indirect response to the presence of inflammation in the organization. In this study, by observing the Kuijieling UC rat colon mucosa of iNOS, MPO activity, its mechanism of action discussed. Grouping and treatment with the foregoing, the colonic mucosa specimens collected after the end of treatment and extraction of whole-cell protein, using biochemical kit was detected in the colonic mucosa of iNOS, MPO activity, and analyzed statistically.The results indicate that the activity of iNOS in NC was4.407±1.448U/mgprot, was significantly higher than in NC group2.530±0.424U/mgprot (P<0.01); the activity of iNOS in KHD, KMD group were3.105±1.121U/mgprot,2.945±0.656U/mgprot, were significantly lower than that in MC group (P<0.05). In addition the activity of MPO in MC group was0.960±0.558units/g wet tablets, was significantly lower than that in NC group0.455±0.365units/g wet tablets(P<0.05); the activity of MPO in KHD, KMD group were0.348±0.194units/g wet tablets,0.513±0.219units/g wet tablets were lower than that in MC group (P<0.01, P<0.05). Tips of KD can reduce UC rat colonic mucosa in iNOS, MPO activity.4Effect of KD on IL-10in Colonic Mucosa of UC Rats ModelCytokines play an important role in the pathogenesis of UC, and progress, pro-inflammatory cytokines and suppression of inflammatory cytokine imbalance is the important factor in development of ulcerative colitis. NF-kB regulates cytokine release in patients with UC, they are closely related to the incidence and progression of UC. In this study, grouping, and treatment with the aforementioned colonic mucosa specimens collected after the end of treatment, using the double antibody sandwich ABC-ELISA detection of colon mucosa of IL-10levels, and statistically analyzed. The results show that the MC group and the content of IL-10in colonic mucosa of10.494±3.410ng/L, was significantly higher than that in NC group3.952±1.277ng/L (P<0.01); KHD group of colonic mucosa in the content of IL-10is16.123±3.607ng/L, was significantly larger than the model group (P<0.01). Tip UC rat colonic mucosa IL-10content increases gradually, while that of KD can improve UC rat colonic mucosa in suppression of inflammatory cytokine IL-10levels.In short, the NF-κB pathway in pathogenesis of UC play a decisive role in the development process, the activation process and mechanism of action and many cytokines and inflammatory mediator related. COX-2, iNOS, MPO are NF-κB activation important molecules, and NF-κB activation were positively correlated, in NF-κB activation in the process of promoting effect. While IL-10for suppression of inflammatory cytokine, can inhibit the NF-κB activation, the process may be through inhibition of IKK activation, block IκB a Degradation, and inhibition of NF-κB activity, reduce the expression of cytokines, the termination of the UC intestinal immune and inflammatory reaction. Clinical studies have demonstrated that COX-2, iNOS, MPO and UC in the pathogenesis of a close relationship, the inflammation of the mucous membrane of overexpression and UC content of colonic mucosal inflammation was positively related to the degree of. While the IL-10has been identified as susceptibility genes of UC. This research selects the clinical experience of KD on TNBS synthesis of rat model of UC, after treatment of colon mucosa in IκB a Protein positive ratio increases significantly, the IKK β protein positive cells was significantly decreased colonic mucosal tissue, the expression of COX-2protein was significantly reduced, colonic mucosa tissue in iNOS, MPO activity decreased the blood, the IL-10content increased significantly. Show of KD can improve UC rat colonic mucosa of IκB α protein expression, reduced IKK β protein expression and has inhibitory effect on COX-2, iNOS, MPO, have stimulative effect on IL-10, which may be the mechanism of the effect of treatment of UC. |
PDF Full Text Request